'Gear up and get ready': Collaborative curriculum for radiographers supporting percutaneous nephrolithotomy in the operating theatre.

INTRODUCTION: Newly qualified radiographers often find working in the operating theatre (OT) challenging and intimidating. These perceptions, which inhibit confidence, may hinder their effectiveness in interprofessional teamwork, which may in turn adversely affect patient outcomes. A collaborative education programme was designed, building upon the foundations of competency-based education (CBE) and simulation-based mastery learning (SBML) to examine its potential in mitigating these perceptions. The objective of this research was to assess participants' experience and level of competency after attending the curated collaborative educational programme. METHODS: The programme was developed based on the Analysis, Design, Development, Implementation, and Evaluation (ADDIE) model and comprises two teaching and learning phases: educational session and simulation. A collaborative approach was undertaken to develop an assessment checklist for the interprofessional simulation. Requirements for the simulation, such as scenario design, information and storyboard, task trainer, logistics, and learners' briefing, debrief, and feedback, were identified and assembled. The radiographers' performance was recorded using a practical skills assessment checklist and a theory assessment. RESULTS: Twelve radiographers participated and showed improvement in their self-rating of learning objectives before and after the programme. The median (interquartile range) score achieved in the theory assessment, out of a possible of 11, was 9.00 (7.75-9.50). The median (interquartile range) score achieved in the simulation component, out of a possible of 16, was 15.00 (14.00-15.00). There was statistically significant difference in self-perceived performance in all learning objective domains. CONCLUSION: The findings from the programme were promising. The use of simulation and an assessment checklist proved to be useful learning tools in preparing newly qualified radiographers for work in the OT. IMPLICATIONS FOR PRACTICE: Assessment checklists are valuable tools that should be considered to facilitate teaching and learning. The use of interprofessional simulation activities can support radiographers in developing knowledge, professional skills, and clinical competency. It should be conducted in a timely manner to facilitate the introduction to role understanding and effective communication.

[Clinical analysis of retropharyngeal lymph node metastasis in patients with hypopharyngeal squamous cell carcinoma].

Objective: To analyze the incidence and the related risk factors of retropharyngeal lymph node metastasis in patients with hypopharyngeal squamous cell carcinoma, evaluate the accuracy of preoperative enhanced CT in judging retropharyngeal lymph node metastasis, and investigate the impact of retropharyngeal lymph node metastasis on the prognosis. Methods: Retrospective analyses were made on 398 patients with hypopharyngeal squamous cell carcinoma who underwent surgery as the primary therapy and accepted retropharyngeal lymph node exploration and clearance during surgery in Shandong Provincial ENT Hospital from January 2014 to December 2019. Multivariate logistic regression analysis was used to clarify the related risk factors of retropharyngeal lymph node metastasis. Multivariate Cox regression analysis was used to investigate the impact of retropharyngeal lymph node metastasis on prognosis. The retropharyngeal lymph nodes of 218 cases with available preoperative enhanced CT images were evaluated by two experienced radiologists and compared with postoperative pathological results. Results: Retropharyngeal lymph node metastasis were confirmed in 54 of 398 (13.6%) cases according to postoperative pathology. The sensitivity and specificity of preoperative enhanced CT in the diagnosis of retropharyngeal lymph node metastasis were 34.6% and 91.1%, respectively, and the overall accuracy was 84.4%. Multivariate logistic regression analysis showed that the site of the primary lesion and pathological N stage were independent risk factors for retropharyngeal lymph node metastasis in hypopharyngeal squamous cell carcinoma. Patients with primary lesion located in the posterior wall of hypopharynx (OR=4.83, 95% CI: 1.27-18.40), N2 stage (OR=6.30, 95% CI: 2.25-17.67), and N3 stage (OR=26.89, 95% CI: 5.76-125.58) were prone to retropharyngeal lymph node metastasis. The 5-year overall survival rate of the 398 patients was 50.4%, and the 5-year disease-free survival rate was 48.3%. Multivariate Cox regression analysis showed that T stage, N stage, retropharyngeal lymph node metastasis, and radiotherapy were independent influencing factors for overall survival (T stage: HR=1.28, 95% CI: 1.06-1.54; N stage: HR=1.26, 95% CI: 1.14-1.40; retropharyngeal lymph node metastasis: HR=2.13, 95% CI: 1.47-3.08; radiotherapy: HR=0.54, 95% CI: 0.38-0.76) and disease-free survival of patients with hypopharyngeal squamous cell carcinoma (T stage: HR=1.26, 95% CI: 1.06-1.51; N stage: HR=1.25, 95% CI: 1.13-1.37; retropharyngeal lymph node metastasis: HR=2.24, 95% CI: 1.56-3.21; radiotherapy: HR=0.55, 95% CI: 0.40-0.77). Conclusions: Metastasis of retropharyngeal lymph nodes in hypopharyngeal squamous cell carcinoma is not rare. Enhanced CT is of low accuracy and limited value in diagnosing retropharyngeal lymph node metastasis. Primary lesions located in the posterior wall of the hypopharyngx, N2 stage, and N3 stage are independent high-risk factors for retropharyngeal lymph node metastasis. The prognosis of hypopharyngeal cancer patients with retropharyngeal lymph node metastasis is worse, and active surgical exploration and clearance can effectively reduce the mortality caused by retropharyngeal lymph node metastasis.

[Exploration on appropriate fetal weight of twin pregnancies at different gestational weeks and pregnancy complications and outcomes].

Objective: To explore the appropriate fetal weight of twin pregnancies at different gestational weeks and the association with pregnancy complications and outcomes. Methods: Fetal weight at different gestational weeks and related pregnancy complications and outcomes from 1 225 twin pregnancies, who gave birth at Peking University First Hospital from January 2004 to December 2020, were analyzed in this study, including hypertensive disorders in pregnancy, gestational diabetes mellitus (GDM), fetal growth restriction (FGR), fetal distress, preterm birth and neonatal asphyxia. The appropriate fetal weight of twin pregnancies at different gestational weeks were analysed based on the information from 616 twin pregnancies without complications (except preterm birth), and were expressed as P10~P90. The chi-square test was used to compare the risk of pregnancy complications and adverse outcomes in large for gestational age (LGA), appropriate for gestational age (AGA) and small for gestational age (SGA) twin pregnancies and the difference in incidence of pregnancy complications and adverse outcomes in different years. Results: The appropriate fetal weights of normal twin pregnancies at 28 to 37 weeks and 38-40 weeks of gestation were 910-1 255 g, 996-1 518 g, 1 105-1 785 g, 1 295-1 825 g, 1 336-2 000 g, 1 754-2 321 g, 1 842-2 591 g, 1 913-2 615 g, 2 150-2 847 g, 2 350-3 130 g and 2 450-3 250 g, respectively. The incidences of hypertensive disorders in pregnancy, FGR, fetal distress and neonatal asphyxia related to SGA twin pregnancies were significantly higher than AGA twin pregnancies (all P<0.05). The incidence of GDM in twin pregnant from 2017 to 2020 was higher than that from 2004 to 2009 or from 2010 to 2016, but the incidence of fetal distress and neonatal asphyxia were lower than those from 2010 to 2016, and the differences were statistically significant (all P<0.05). Conclusions: The appropriate weights of twin fetuses at different gestational weeks are different from singleton. The incidence of pregnancy complications and adverse outcomes in AGA fetuses is significantly lower than that in SGA fetuses under the specific weight standard for twin fetuses, which could provide a practical basis for clinical management of twin pregnancy.

[Comparison of four-coagulation-tests values in normal pregnant women during early and late pregnancy and the influence of age].

Objective: To explore and compare the reference ranges of four coagulation tests in normal pregnant women during early and late pregnancy and the influence of age. Methods: Values of four coagulation tests from 4 974 pregnant women, who gave single birth at Peking University First Hospital, Obstetrics and Gynecology Hospital of Fudan University, West China Second University Hospital, Peking University Third Hospital and Shengjing Hospital of China Medical University from February 2017 to July 2020, were measured and analyzed in this study, including prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib) and thrombin time (TT). The four normal reference ranges of coagulation during early and late pregnancy phases were expressed as P2.5-P97.5. The difference of two pregnancy phases was compared by non-parametric test of two related samples. And the difference between pregnant women of advanced and non-advanced age in the same pregnancy phase was compared by independent sample non-parametric test. Chi-square test was used to compare the incidence of pregnancy complications in different coagulation reference ranges. Results: The reference ranges of PT of normal pregnant women's early and late pregnancy were 10.0-13.9 s and 9.6-12.3 s, the reference ranges of APTT were 22.6-35.3 s and 22.4-30.9 s, the reference ranges of Fib were 2.4-5.0 g/L and 3.0-5.7 g/L, the reference ranges of TT were 12.0-19.0 s and 11.5-18.4 s. Compared with early pregnancy, PT, APTT and TT shortened significantly, while the Fib significantly increased in late pregnancy (all P<0.001). PT, APTT and TT of advanced and non-advanced age pregnant women were significantly different (all P<0.01). Compared with the ranges of non-pregnant population, more pregnant women were included in the normal pregnant reference ranges of PT in early pregnancy and APTT in the early and late pregnancy, while the incidence of pregnancy complications had no significant differences (all P>0.05). The incidence of fetal distress was higher and the incidence of preterm birth was lower in the reference range of PT in late pregnancy. The incidence of gestational diabetes mellitus was higher in the early and late gestational Fib reference ranges, and the incidence of hypertensive disorders in pregnancy was higher in the late gestational Fib reference range (all P<0.05). Conclusions: The coagulation function of pregnant women increases significantly with the growth of pregnancy, and there is a significant difference between advanced significantly and non-advanced age pregnant women. The recommended ranges of normal pregnant women's early and late pregnancy PT are 10.0-13.9 s and 9.6-12.3 s, the recommended ranges of APTT are 22.6-35.3 s and 22.4-30.9 s, the recommended ranges of TT are 12.0-19.0 s and 11.5-18.4 s. The appropriate ranges of normal pregnant women's early and late pregnancy Fib still need further exploration.

[The effect of cervical lymph node metastasis density on prognosis of hypopharyngeal carcinoma].

Objective: To investigate the relationship between the cervical lymph node density (LND) and the prognosis of hypopharyngeal carcinoma. Methods: The clinical and pathological data of 241 patients with hypopharyngeal carcinoma who underwent surgery in Shandong Provincial ENT Hospital from January 2014 to December 2017 were retrospectively analyzed, including 229 males and 12 females, aged 37-81 years. The LND was calculated, i.e. the ratio of the number of lymph nodes with metastasis to the total number of lymph nodes removed. The patients were divided into low LND group and high LND group based on the cutoff value of LND determined by receiver operating characteristic curve (ROC curve). The univariate and multivariate analyses of the disease-free survival (DFS) and the overall survival (OS) were performed in two groups. Results: With the cutoff value of 0.068, 165 patients were in the low LND group (<0.068) and 76 patients in the high LND group (≥0.068). T stage, N stage, maximum lymph node diameter, extracellular invasion of lymph node, and postpharyngeal lymph node metastasis were associated with LND (statistical values were -3.15, -6.82, 23.37, 20.44, and 30.18, respectively, all P values were<0.05). The univariate analysis showed that age, T stage, N stage, maximum diameter of cervical lymph nodes, extracapsular invasion, retropharyngeal lymph node metastasis and LND were the main factors affecting the patients' DFS (χ2=9.31, 7.30, 20.09, 15.30, 9.04, 19.44, 50.27, all P values<0.05) and OS (χ2 were 5.02, 12.94, 18.28, 15.91, 7.95, 16.88, 49.45, all P values<0.05). Multivariate analysis showed that patients with age≤60 years old and LND≥0.068 had reduced DFS [HR values were 0.61 (95%CI 0.43-0.88) and 2.23 (95%CI 1.44-3.45), both P values<0.05]; patients with advanced T stage and LND≥0.068 had reduced OS [HR values were 1.73 (95%CI 1.02-2.93) and 2.39 (95%CI 1.51-3.80), both P values<0.05]. Conclusion: LND is a prognostic factor for patients with hypopharyngeal carcinoma after surgery, with worse prognosis in patients with LND≥0.068.

[Establishment and preliminary application of a recombinase-aided isothermal amplification assay-based multiplex nucleic acid assay for detection of three Echinococcus species].

OBJECTIVE: To establish a multiplex nucleic acid assay for rapid detection of Echinococcus multilocularis, E. granulosus and E. shiquicus based on the recombinase-aided isothermal amplification assay (RAA) and to preliminarily assess its diagnostic efficiency. METHODS: The mitochondrial genomic sequences of E. multilocularis (GenBank accession number: NC_000928), E. granulosus (GenBank accession number: NC_044548) and E. shiquicus (GenBank accession number: NC_009460) were used as target sequences, and three pairs of primers were designed based on the RAA primer design principle and synthesized for the subsequent multiple RAA amplification. The genomic DNA of E. multilocularis, E. granulosus and E. shiquicus at different concentrations and the recombinant plasmids containing the target gene at various concentrations were amplified to evaluate the diagnostic sensitivity of the multiplex RAA assay, and the genomic DNA of E. multilocularis, E. granulosus, E. shiquicus, Taenia multiceps, T. saginata, T. asiatica, Dipylidium caninum, T. hydatigena, Toxocara canis, Fasciola hepatica, T. pisiformis, Mesocestoides lineatus and Cryptosporidiumn canis was detected using the multiplex RAA assay to evaluate its specificity. In addition, the reaction condition of the multiplex RAA assay was optimized, and was then employed to detect the tissues with echinococcosis lesions, simulated canine fecal samples and field captured fox fecal samples to examine its application values. RESULTS: The multiplex RAA assay was effective to specifically amplify the mitochondrial gene fragments of E. multilocularis, E. granulosus and E. shiquicus within 40 min at 39 °C, with sequence lengths of 540, 430 bp and 200 bp, respectively. This multiplex RAA assay showed the minimum detection limits of 2.0, 2.5 pg/µL and 3.1 pg/µL for detection of the genomic DNA of E. multilocularis, E. granulosus and E. shiquicus, and presented the minimum detection limit of 200 copies/µL for detection of the recombinant plasmids containing E. multilocularis, E. granulosus and E. shiquicus target genes. This multiplex RAA assay was effective to simultaneously detect single and multiple infections with E. multilocularis, E. granulosus and E. shiquicus, but failed to amplify the genomic DNA of T. multiceps, T. saginata, T. asiatica, D. caninum, T. hydatigena, T. canis, F. hepatica, T. pisiformis, M. lineatus and C. canis. In addition, the optimized multiplex RAA assay was effective to detect all positive samples from the tissue samples with echinococcosis lesions, simulated canine fecal samples and field captured fox fecal samples, which was fully consistent with the detection of the single PCR assay. CONCLUSIONS: A sensitive and specific multiplex nucleic acid assay for rapid detection of E. multilocularis, E. granulosus and E. shiquicus has been successfully established.

[Effects of interpregnancy interval on pregnancy outcomes of subsequent pregnancy: a multicenter retrospective study].

Objective: To explore the effects of interpregnancy interval (IPI) on pregnancy outcomes of subsequent pregnancy. Methods: A multicenter retrospective study was conducted in 21 hospitals in China. Information of age, height, pre-pregnancy weight, IPI, history of diseases, complications of pregnancy, gestational age of delivery, delivery mode, and pregnancy outcomes of the participants were collected by consulting medical records of pregnant women who had two consecutive deliveries in the same hospital during 2011 to 2018. The participants were divided into 4 groups according to IPI:<18 months, 18-23 months, 24-59 months and ≥60 months. According to the WHO's recommendation, with the IPI of 24-59 months group as a reference, to the effects of IPI on pregnancy outcomes of subsequent pregnancy were analyzed. Stratified analysis was further carried out based on age, history of gestational diabetes mellitus (GDM), macrosomia, and premature delivery, to explore the differences in the effects of IPI on pregnancy outcomes among women with different characteristics. Results: A total of 8 026 women were included in this study. There were 423, 623, 5 512 and 1 468 participants in <18 months group, 18-23 months group, 24-59 months group and ≥60 months group, respectively. (1) The age, pre-pregnancy body mass index (BMI), history of cesarean section, GDM, gestational hypertension and cesarean section delivery rate of <18 months group, 18-23 months group, 24-59 months group and ≥60 months group were gradually increased, and the differences were statistically significant (P<0.05). (2) After adjusting for potential confounding factors, compared with women in the IPI of 24-59 months group, the risk of premature delivery, premature rupture of membranes, and oligohydramnios were increased by 42% (OR=1.42, 95%CI: 1.07-1.88, P=0.015), 46% (OR=1.46, 95%CI: 1.13-1.88, P=0.004), and 64% (OR=1.64, 95%CI: 1.13-2.38, P=0.009) respectively for women in the IPI≥60 months group. No effects of IPI on other pregnancy outcomes were found in this study (P>0.05). (3) After stratified by age and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would significantly increase the risk of oligohydramnios for women with advanced age (OR=2.87, 95%CI: 1.41-5.83, P=0.004); and <18 months could increase the risk of premature rupture of membranes for women under the age of 35 (OR=1.59, 95%CI: 1.04-2.43, P=0.032). Both the risk of premature rupture of membranes (OR=1.58, 95%CI: 1.18-2.13, P=0.002) and premature delivery (OR=1.52, 95%CI: 1.07-2.17, P=0.020) were significantly increased in the IPI≥60 months group. After stratified by history of GDM and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would lead to an increased risk of postpartum hemorrhage for women with a history of GDM (OR=5.34, 95%CI: 1.45-19.70, P=0.012) and an increased risk of premature rupture of membranes for women without a history of GDM (OR=1.44, 95%CI: 1.10-1.90, P=0.009). After stratified by history of macrosomia and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months could increase the proportion of cesarean section for women with a history of macrosomia (OR=4.11, 95%CI: 1.18-14.27, P=0.026) and the risk of premature rupture of membranes for women without a history of macrosomia (OR=1.46, 95%CI: 1.12-1.89, P=0.005). After stratified by history of premature delivery and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would significantly increase the risk of premature rupture of membranes for women without a history of premature delivery (OR=1.47, 95%CI: 1.13-1.92, P=0.004). Conclusions: Both IPI≥60 months and <18 months would increase the risk of adverse pregnancy outcomes in the subsequent pregnancy. Healthcare education and consultation should be conducted for women of reproductive age to maintain an appropriate IPI when they plan to pregnant again, to reduce the risk of adverse pregnancy outcomes in the subsequent pregnancy.

[Effect of glucose on Akt2 mRNA and methylation level and the association between early-pregnancy Akt2 mRNA level in peripheral blood and pre-pregnancy BMI and GDM].

Objective: To explore the effect of glucose on Akt2 gene methylation and mRNA level in the trophoblast cell line HTR8/SVneo and the association between first-trimester Akt2 mRNA level in peripheral blood and pre-pregnancy body mass index (BMI) and gestational diabetes mellitus (GDM). Methods: The cell model of intrauterine hyperglycemia in pregnant women with GDM was established, and divided into 5 groups based on the different glucose concentrations (2.5, 5.0, 10.0, 25.0, and 40.0 mmol/L). The mRNA levels of Akt2 gene were detected by real-time quantitative PCR, and the methylation levels of Akt2 gene promoter region were detected by mass spectrometry. According to pre-pregnancy BMI and diagnosis of GDM, 60 pregnant women who had given birth at Peking University First Hospital during December 2014 to July 2016 were classified into overweight non-GDM group, overweight GDM group, obese non-GDM group and obese GDM group. Real-time quantitative PCR analyses were used to detect the levels of Akt2 mRNA in first-trimester peripheral blood of the 4 groups. Results: (1) Level of Akt2 mRNA significantly rose with the glucose concentration of medium increased, showing concentration dependency (all P<0.05). Compared with the 25.0 mmol/L group, the following methylation levels changed significantly (all P<0.05): cytosine-phosphate-guanine (CpG) 10, CpG23, and CpG24.5 of Akt2 gene promoter region in the 5.0 mmol/L group, CpG23 and CpG24.5 in the 2.5 mmol/L group, and CpG10 in the 10.0 mmol/L group. Compared with the 5.0 mmol/L group, CpG10 in the 40.0 mmol/L group showed methylation change (P<0.05). (2) Compared with the overweight non-GDM group [1.04(0.90~1.26)], overweight GDM group [2.10(1.85~2.28)] and obese GDM group [1.68(0.82~2.43)] all had higher Akt2 mRNA levels and the differences were statistically significant (all P<0.05). Compared to obese GDM group [1.68(0.82~2.43)], the level of Akt2 mRNA was higher in overweight GDM group [2.10(1.85~2.28)] and lower in obese non-GDM group [1.00(0.71~2.17)], but the differences were not statistically significant (all P>0.05). Conclusions: Glucose might affect Akt2 mRNA level by changing the methylation of Akt2 gene promoter region, and the change might appear in the first trimester of pregnant women with GDM, especially for women with lower pre-pregnancy BMI.

Effects of main cereal type and feed form on production performance, egg quality and egg sanitary indices of laying hens.

1. The experiment was conducted to evaluate the effects of cereal types (maize or wheat) and feed forms (pelleted or mash feed) on production performance, egg quality and egg sanitary indices in laying hens.2. Three hundred and sixty hens (Jinghong No. 1) at 18 weeks of age were randomly assigned to four treatments with six replicates of 15 hens per replicate according to a 2 × 2 factorial design with two cereal types (maize or wheat) and two feed forms (pelleted or mash feed).3. Compared with the wheat-based diet, the maize-based diet improved (P < 0.05) average egg weight of laying hens. Yolk colour of hens fed with the maize-based diet was higher (P < 0.05) in comparison to those fed the wheat-based diet, while Haugh units were lower (P < 0.05) for the maize-based treatment. Egg mass and average daily feed intake of hens fed the pelleted diet were higher (P < 0.05) than of those fed the mash diet. However, the mash diet improved (P < 0.05) yolk colour compared with the pelleted diet. The percentage of dirty eggs for the wheat-based diet was higher (P < 0.05) than for the maize-based diet. The percentage of dirty eggs was higher (P < 0.05) in birds fed the pelleted diet compared with those fed the mash diet. There were interactions (P < 0.05) between cereal type and feed form, with regard to average egg weight and shell thickness.4. In conclusion, dietary cereal type affected average egg weight, yolk colour and Haugh units in eggs, while feed form influenced egg mass, average daily feed intake and yolk colour.

[Spatial-temporal cluster of hemorrhagic fever with renal syndrome in Hebei province, 2005-2016].

Objective: To explore the spatial-temporal distribution and epidemic characteristics of hemorrhagic fever with renal syndrome (HFRS) in Hebei province from 2005 to 2016. Methods: Records of HFRS cases reported from each county in Hebei during January 2005 to December 2016 were collected from National Notifiable Disease Surveillance System (NNDSS). Global and local spatial association statistics were used to measure the spatial autocorrelation and software GeoDa 1.2.0. Software SaTScan 9.4.1 was used to analyze spatiotemporal clusters. Software ArcGIS 10.2 was used to visualize the yearly scan results. Results: In Hebei province, a total of 8 437 human HFRS cases reported from 170 counties with an annual incidence rate of 0.99/100 000 population during 2005-2016. The peak incidence season was spring. Global spatial autocorrelation analysis on the incidence of HFRS at county-level showed that the value of Moran's I were all above 0 (P<0.05), indicating that the significant spatial cluster. The result of local indicators on spatial association (LISA) analysis revealed that identified hot spots were mainly in northeastern area, while cold spots were found in some counties of central and southern areas. Spatial-temporal scan detected that the primary cluster of HFRS incidence was mainly distributed in Qinhuangdao city and Tangshan city, including 11 counties (city/district): Beidaihe district, Haigang district, Funing district, Shanhaiguan district, Changli county, Lulong county and Qinglong Manchu autonomous county in Qinhuangdao city, and Qian'an city, Laoting county, Luanzhou city and Luannan county in Tangshan city (RR=39.64, P<0.001), during January-July in 2005. Conclusions: There were significant spatial-temporal cluster of HFRS in Hebei from 2005 to 2016. The cluster areas of HFRS were mainly in northeastern Hebei, it is necessary to strengthen the prevention and control programs of HFRS in these areas.

Comparison of Mitochondrial Function in Boar and Bull Spermatozoa Throughout Cryopreservation Based on JC-1 Staining.

BACKGROUND: Poor reproductivity hampers the commercialization of cryopreserved boar semen. OBJECTIVE: This study was to determine the differences in the sperm mitochondrial function between boar and bull semen at different cryopreservation stages. MATERIALS AND METHODS: Boar and bull fresh, equilibrated, and frozen-thawed spermatozoa were evaluated for mitochondrial function using JC-1 under a fluorescent microscope. RESULTS: Bull and boar percentage of spermatozoa staining green (PSSG) showed no difference between fresh and equilibrated semen (P> 0.05). However, frozen-thawed bull and boar semen demonstrated significantly higher PSSG (P < 0.01) than fresh and equilibrated semen. Frozen-thawed boar semen represented a significantly higher PSSG (P < 0.01) than bull semen. CONCLUSION: Negative cryopreservation influence on boar and bull spermatozoa was not significantly produced by pre-freezing procedures, but rather by freezing and thawing. Cryopreservation has more pronounced negative effects on boar than on bull spermatozoa, which partly explains lagged commercialization of frozen boar semen.

[Characteristics of pre-gestational diabetes mellitus diagnosed during pregnancy and the effects on pregnancy outcomes].

Objective: To analyze the characteristics of pre-gestational diabetes mellitus (PGDM) diagnosed during pregnancy (missed diagnosis before pregnancy), and to evaluate the effects of diagnostic time on pregnancy outcomes. Methods: A retrospective study of 746 pregnant women who were diagnosed PGDM and delivered in Peking University First Hospital from January 1st, 2005 to December 31st, 2015 was conducted. The patients were divided into 2 group. Those diagnosed PGDM before pregnancy were defined as Group diagnosed before pregnancy, and those diagnosed during pregnancy were defined as Group diagnosed during pregnancy. In Group diagnosed during pregnancy, those diagnosed before 24 gestational weeks were defined as Group diagnosed during pregnancy A, and those diagnosed after 24 weeks were defined as Group diagnosed during pregnancy B. The prevalence of adverse pregnancy outcomes in each group were analyzed. Results: (1) Rate of missed diagnosis for PGDM: the incidence of PGDM diagnosed before pregnancy was 32.2% (240/746), and those diagnosed during pregnancy (missed diagnosis before pregnancy) was 67.8% (506/746). (2) Blood glucose control during pregnancy: ①Group diagnosed before pregnancy and Group diagnosed during pregnancy: the highest glycosylated hemoglobin (HbA1c) in Group diagnosed before pregnancy was (6.6±1.1)%, higher than that in Group diagnosed during pregnancy [(6.3±1.0)%, P=0.019]. However, there was no significant difference in the average HbA1c level between the 2 groups (P=0.616). The insulin needed percentage [90.8%(218/240) vs. 53.8%(272/506)] in Group diagnosed before pregnancy were higher than that in Group diagnosed during pregnancy (P<0.01). ②Group diagnosed during pregnancy A and B: the highest HbA1c in Group diagnosed during pregnancy A was (6.9±1.3)%, higher than that in Group diagnosed during pregnancy B [(6.1±0.8)%, P<0.05]. And the average HbA1c in Group diagnosed during pregnancy A [(6.4±0.8)%] was also higher than that in Group diagnosed during pregnancy B [(6.0±0.8)%, P<0.05]. In Group diagnosed during pregnancy B, 46.1% (187/406) used insulin, lower than the percentage in Group diagnosed during pregnancy A (85.0%, 85/100; P<0.01). ③There were no significant differences in the highest HbA1c and the average HbA1c between Group diagnosed during pregnancy A and Group diagnosed before pregnancy (P=0.020, P=0.037). There was neither no significant difference in the percentage used insulin during pregnancy between them (P=0.128). There were significant differences in the highest HbA1c and the average HbA1c between Group diagnosed during pregnancy B and Group diagnosed before pregnancy (P<0.01, P=0.014). There was also significant difference in the percentage used insulin during pregnancy between them (P<0.01). (3) Pregnancy outcome: ①Group diagnosed before pregnancy and Group diagnosed during pregnancy: the cesarean section rate [72.5% (174/240) vs. 59.7% (302/506)] in Group diagnosed before pregnancy were higher than those in Group diagnosed during pregnancy (P<0.01). However, there were no significant differences in preterm birth rate, pre-eclampsia, macrosomia percentage, percentage of neonates being hospitalized between the 2 groups (P=0.546, P=1.000, P=0.671, P=0.804) . ②There was no significant difference in preterm birth rate,cesarean delivery rate, macrosomia percentage, pre-eclampsia rate, percentage of neonates being hospitalized between Group diagnosed during pregnancy A and Group diagnosed during pregnancy B (P=0.887, P=0.495, P=0.841, P=1.000, P=1.000).③There was no significant difference in preterm birth rate, cesarean delivery rate, macrosomia percentage, pre-eclampsia rate, percentage of neonates being hospitalized between Group diagnosed during pregnancy A and Group diagnosed before pregnancy (P=0.875, P=0.093, P=0.662, P=1.000, P=0.837). The cesarean delivery rate was lower in Group diagnosed during pregnancy B than that in Group diagnosed before pregnancy (P=0.001). However, there were no significant differences in preterm birth rate, macrosomia percentage, pre-eclampsia rate, percentage of neonates being hospitalized between them (P=0.530, P=0.776, P=1.000, P=0.797). Conclusions: The diagnosis of PGDM is commonly missed before pregnancy. Fasting plasma glucose should be used as screening test to identify PGDM at pre-pregnancy examination or first antenatal care. Using abnormal value of 2-hour glucose after 24 gestational weeks as the only way to diagnose PGDM is not suitable.

Second harmonic generation from an individual amorphous selenium nanosphere.

Among the numerous nonlinear optics effects, second harmonic generation (SHG) is always a hotspot and it is extensively used for optical frequency conversion, biomedical imaging, etc. However, SHG is forbidden in a medium with inversion symmetry under the electric-dipole approximation. Here, we demonstrated SHG from a single amorphous selenium (a-Se) nanosphere under near-infrared femtosecond pulse excitation. It was found that SH spectra are tunable with the size of a-Se nanospheres and the SHG efficiency of a single a-Se sphere with a diameter over 300 nm is estimated at 10(-8). We also established two physical mechanisms of SHG from the amorphous nanospheres. There is an electric-dipole contribution to the second-order nonlinearity in view of the inevitable structural discontinuity at the surface. The discontinuity of the normal component of the electric field strength leads to the quadrupole-type contributions arising from the large electric field gradient. The SHG process can be enhanced by resonance near the fundamental wavelength, giving rise to the detectable second harmonic (SH) spectra of a single a-Se nanosphere (d > 300 nm) or two small a-Se nanospheres (d = 200 nm) aggregated into a dimer, while the single nanosphere with smaller size (d > 300 nm) is undetectable. As an essential trace element for animals, a-Se features unique biological compatibility and has specific properties of optical nonlinearity within the optical window in biological tissue. This discovery makes a-Se nanospheres promising both in nonlinear optics and biomedicine.

[Incidence of singleton macrosomia in Beijing and its risk factors].

OBJECTIVE: To evaluate the prevalence of macrosomia in Beijing in 2013 and identify its risk factors. METHODS: Retrospective six months analysis of 14 188 full-term singleton pregnant women from 15 hospitals with different levels in Beijing in 2013. Each participant's demographic data and medical information were collected individually by questionnaires. Multiple logistic regression analysis was used to examine the associations between variables and the risk of macrosomia. RESULTS: (1)The total prevalence of macrosomia was 7.069% (1 003/14 188) in Beijing in 2013. (2)The prevalence varied between the 15 hospitals, the lowest was 5.36% (89/1 659), while the highest reached 8.80%(46/523). Furthermore, the incidence of macrosomia was 1.284 times (95% CI: 1.114- 1.480, P=0.001) higher in the second graded hospitals than that in the tertiary hospitals. (3) Multiple logistic regression analyses showed that risk factors for macrosomia were maternal height≥160 cm (adjusted OR=1.875, 95% CI: 1.559- 2.256), pre-pregnant body mass index (p-BMI) ≥24.0 kg/m(2) (24.0- 27.9 kg/m(2): adjusted OR=1.696, 95% CI: 1.426- 2.018; p-BMI≥28.0 kg/m(2): adjusted OR=2.393, 95% CI: 1.831- 3.127), gestational weight gain (GWG) ≥15.9 kg (adjusted OR=2.462, 95% CI: 2.125- 2.853), gravidity>1 (adjusted OR=1.408, 95% CI: 1.224- 1.620), gestational weeks≥40 (adjusted OR=2.007, 95%CI: 1.745-2.308) and gestational diabetes mellitus (adjusted OR=1.522, 95%CI: 1.298-1.784). GWG≥15.9 kg, p-BMI≥28.0 kg/m(2) and gestational weeks≥40 were three risk factors that had the strongest associations with macrosomia (all P<0.01). CONCLUSIONS: The prevalence of macrosomia in hospitals with different levels is obvious different. Gestational weeks, p-BMI and GWG are three main controllable risk factors for macrosomia, thus should receive more attentions.

Identification of severe gestational diabetes mellitus after new criteria used in China.

OBJECTIVE: To evaluate the way to identify severe gestational diabetes mellitus (GDM) according to the Ministry of Health (MOH) criteria associated with high risk of adverse pregnancy outcomes. STUDY DESIGN: Medical records of 9803 pregnant women attending Peking University First Hospital were analyzed retrospectively. Participants diagnosed as GDM were divided into different groups according to the different number of oral glucose tolerance test (OGTT) results and the prepregnancy body mass index (BMI). Participants without GDM were included in group N. The incidence of adverse pregnancy outcomes was analyzed according to incremental differences in prepregnancy BMI and the number of abnormal OGTT result. RESULT: (1) There were 21.8% (2133/9803) women diagnosed as GDM. (2) The frequency of large for gestational age in group 2 (GDM with 2 or more abnormal OGTT value; 21.6%) was significantly higher than group 1 (GDM with only 1 abnormal OGTT value; 16.8%) and group N (13.2%), and there was also significant difference between group 1 and group N. (3) Risk of adverse pregnancy outcomes in GDM would be increased in women with prepregnancy body mass index ⩾24 kg m(-2) in GDM. (4) Women with BMI <24 kg m(-2) in group 1 have low risk of adverse pregnancy outcomes and seldom need insulin (1.3%) in pregnancy. (5) There were 1142 cases (53.5%) of severe GDM in this study and 991 cases (46.5%) of mild GDM. CONCLUSION: GDM women with ⩾2 abnormal OGTT values or only 1 abnormal OGTT value but with prepregnancy BMI ⩾24 kg m(-2) should be recognized as severe GDM. We should pay more attention to and treat selectively with the severe GDM, especially in rural areas.

Effectiveness of a 95 SNP panel for the screening of breed label fraud in the Chinese meat market.

Breed assignment has proved to be useful to control meat trade and protect the value of special productions. Meat-related frauds have been detected in China; therefore, 95 SNPs selected from the ISAG core panel were evaluated to develop an automated and technologically updated tool to screen breed label fraud in the Chinese meat market. A total of 271 animals from four Chinese yellow cattle (CYC) populations, six Bos taurus breeds, two Bos indicus and one composite were used. The allocation test distinguished European, Japanese and Zebu breeds, and two Chinese genetic components. It correctly allocated Japanese Black, Zebu and British breeds in 100, 90 and 89% of samples, respectively. CYC evidenced the Zebu, Holstein and Limousin introgression. The test did not detect CYC components in any of the 25 samples from Argentinean butchers. The method could be useful to certify Angus, Hereford and Japanese Black meat, but a modification in the panel would be needed to differentiate other breeds.

Pattern of lymph node metastases and its implication in radiotherapeutic clinical target volume in patients with non-small-cell lung cancer: a study of 2062 cases.

OBJECTIVE: To study the pattern of lymph node metastasis (LNM) of non-small-cell lung cancer (NSCLC) and to clarify which node level should be included while undergoing radiotherapy (RT). METHODS: A total of 2062 patients with NSCLC patients who had undergone thoracotomy were retrospectively examined. The clinicopathological factors related to LNM were analysed. RESULTS: The LNM rates (the number of node-positive patients/the total number of patients) in patients with primary tumours in different lobes (left upper lobe, left lower lobe, right upper lobe, right middle lobe and right lower lobe) were 53.25%, 53.87%, 53.77%, 64.67% and 61.58%, respectively. We have found that in all of the clinicopathological factors, including sex, age, tumour location, histological type, maximum diameter, T stage, degree of differentiation and tumour growth pattern, only maximum diameter (p = 0.336) and histological type (p = 0.360) did not have significant correlation with LNM rate. All of the above factors except tumour growth pattern (p = 0.239) and maximum diameter (p = 0.613) were significantly associated with lymph node ratio [LNR, ratio between metastatic and examined lymph nodes (LNs)] in linear regression. CONCLUSION: For patients with NSCLC, LNM rate and LNR can be recommended as applicable parameters for LN involvement. Multiple clinicopathological factors should be considered comprehensively to design the clinical target volume for RT of NSCLC. ADVANCES IN KNOWLEDGE: This article can provide evidence to radio-oncologists how to choose range of lymph nodal clinical target volume when they are treating inoperable patients with NSCLC patients by analysing data of patients after surgery.

Foreign meat identification by DNA breed assignment for the Chinese market.

Methods for individual identification are usually employed for traceability, whereas breed identification is useful to detect commercial frauds. In this study, Chinese Yellow Cattle (CYC) samples plus data from six Bos taurus breeds, two Bos indicus breeds, and one composite breed were used to develop an allocation test based on 22 microsatellites. The test allowed discriminating all foreign breeds from the CYC, although some CYC individuals were wrongly allocated as Limousin or Holstein, probably due to the recent introduction of these breeds into China. In addition, CYC evidenced a previously reported Zebu cline (south-north) and a possible structure within the B. taurus component that should be confirmed. An independent test performed with meat samples of unknown breed origin from Argentina allocated 92% of them to either Angus, Hereford, or their crossbreed, but none was identified as CYC. We conclude that the test is a suitable tool to certify meat of foreign breed origin and to detect adulterations of CYC beef labeled as imported meat.

First Report on the Occurrence of Cucumber mosaic virus on Fragaria ananassa in China.

More than 20 viruses are known to infect strawberry (Fragaria ananassa), and a substantial number of these include new viruses identified since 2000 that can contribute to disease complexes (2). The most serious virus related losses in commercial strawberries are caused by aphid transmitted viruses (3,4,5). A survey was undertaken from 2012 to 2013 to investigate virus prevalence in commercial strawberries in rural areas of Hebei Province around Beijing, China, that were exhibiting virus symptoms. Visual observations revealed that the incidence of virus-like symptoms ranged from 30 to 50% of the plants and these symptoms included yellowing, leaf malformation, sometimes combined with severe stunting and deformed flowers or fruits. Leaf samples were tested for Strawberry vein banding virus (SVBV), Strawberry mottle virus (SMoV), Strawberry mild yellow edge virus (SMYEV), and Strawberry crinkle virus (SCV), which are the four most prevalent aphid-transmitted viruses in single or mixed infections (2). Testing was conducted by RT-PCR using total RNA extracted from fresh symptomatic strawberry leaves (3). SVBV was detected in 58 of 190 samples, but all of the samples tested negative for SMoV, SMYEV, and SCV. Aphids were present on many of the plants, so the samples were tested for Cucumber mosaic virus (CMV) because CMV is prevalent in Beijing gardens and farms, and recently had been shown to infect maize in China (5). This RT-PCR was carried out with the CMV primer pair CM420-F (5'-TGATTCTACCGTGTGGGTGA-3') and CM420-R (5'-CCGTAAGCTGGATGGACAAC-3') to amplify a portion of the capsid protein coding region and the conserved 3'non-translated regions of the genomic RNAs. This test revealed the presence of 43 CMV-positives out of 190 samples, and only 16 of these samples were co-infected with both SVBV and CMV. Samples infected with CMV only had leaf malformations and yellowing, while no CMV was found in symptomless samples. One of the amplified, CMV-specific DNA fragments was sequenced directly from the PCR product and showed 93.8% nucleotide sequence identity and 100% amino acid sequence identity to the CMV subgroup I (GenBank Accession No. D10538) (1). Subsequent ELISA tests for the CMV presence verified the RT-PCR results (Agdia, Elkhart, IN), and transmission electron microscopy observations revealed 28 nm spherical particles characteristic of CMV in strawberry samples tested positive for CMV. However, we were unable to detect either CMV or SVBV in 89 of the 169 samples from symptomatic plants, which suggested possible presence of the other pathogen(s). To the best of our knowledge, this is the first report of natural infections of CMV in strawberry plants. These data suggests that CMV is a potential threat to strawberry production. References: (1) M. Q. K. Andrew et al. Virus taxonomy: IXth Report of the ICTV, 970, Elsevier, 2012. (2) R. R. Martin and I. E. Tzanetakis. Plant Dis. 97:1358, 2013. (3) J. R. Thompson et al. J. Virol. Methods 111:85, 2003. (4) I. E. Tzanetakis et al. Plant Dis. 90:1343, 2006. (5) R. Wang et al. J. Phytopathol. 161: 880, 2013.

Aberrant gene expression profiles in hepatocellular carcinoma detected by microdissection.

The aim of this study was to identify genomic aberrations in hepatocellular carcinoma (HCC) by using laser capture microdissection (LCM) combined with microarray analysis. Samples were procured by LCM from HCC and patient-matched normal liver tissue surgically resected from 4 patients. RNA was isolated from the samples and reverse transcribed into cDNA. After 2-cycle linear amplification and 2-color fluorescent labeling, the cRNA was hybridized onto a whole genome microarray. All genes expressed in the normal and HCC samples were counted and analyzed. Differentially expressed genes were identified and the top 10 up and downregulated genes (totally 20 genes) were further evaluated. LCM was able to accurately capture 50-200 cells from HCC and control tissues. The microarray spectrum showed satisfactory detection of HCC-enriched genes. A total of 1361 differentially expressed genes were identified, among which, 607 were upregulated and 754 were downregulated. Among the top 20 up and downregulated genes, 4 genes had not been documented in the literature as being differentially expressed in any tumors. Thus, LCM is an effective approach for identifying aberrantly expressed genes in HCC, and may lead to the discovery of biomarkers for diagnostic and therapeutic applications.